TWI620511B - Method of producing high purity d-psicose - Google Patents

Abstract

A method for producing D-psicose is disclosed herein. The method for producing D-psicose includes subjecting D-fructose to D-psicose epimerization to produce a D-psicose-containing solution, The solution is subjected to a first cooling and ion purification, so that the purified D-psicose-containing solution is subjected to a first concentration and a second cooling, so that the D-psicose-containing solution that has been subjected to the first concentration and a second cooling is The solution is subjected to chromatography to obtain a mother liquor containing D-fructose and a separation solution containing D-psicose; and subjecting the separation solution containing D-psicose to a second concentration and a third cooling to obtain D- Doseptose crystals, wherein the D-fructose-containing mother liquor produced by chromatography is reused in the D-psicose epimerization.

Description

Method for producing high-purity D-psicose

The present invention relates to a method for manufacturing D-psicose, and more particularly, a method for manufacturing high-purity D-psicose with high yield without thermal deformation during manufacturing.

D-psicose has been reported to be a sweetener different from fructose or sucrose, which is hardly metabolized in the body and has essentially no calories, and hardly affects weight gain due to its inhibition of body fat formation (Matterso T. (. Matsuo, T) and other Asia-Pacific Journal of clinical Untr (Asia Pac.J.Clin.Untr.), 10,233-237,2001; .. (. Matsuo, T) Mate Suo T. and K. Mori Izumo (K Izumori), Asia-Pacific Journal of Clinical Nutrition ( Asia Pac. J. Clin. Nutr. , 13, S127, 2004).

Recently, the present inventors have reported an economical process by isomerizing D-glucose to D-fructose, and then reacting the obtained D-fructose with an immobilized cell capable of producing D-psicose epimerase Method for producing D-psicose (Korean Patent Application No. 10-2009-0118465).

Because the D-psicose-containing reaction solution produced by the enzymatic reaction contains solid A low purity product of D-psicose having a body content of about 20% (w / w) to about 30% (w / w), so D-psicose must be purified by continuous chromatography to produce a purity of 98 % (w / w) or greater than 98% (w / w) high purity D-psicose crystals.

Since the conversion rate of the enzymatic reaction of D-psicose produced from D-fructose by the above method is 20% (w / w) to 30% (w / w), the reaction shows a process flow, wherein the process The amount of mother liquor produced in the process, that is, the amount of D-fructose mother liquor separated from continuous chromatography and the amount of mother liquor separated from crystallization is higher than the amount of D-psicose produced. Therefore, a decrease in the production amount of D-psicose leads to an increase in manufacturing costs, which in turn increases production costs, which causes problems in view of uneconomical industrial production.

Therefore, in order to suppress an increase in manufacturing costs due to low yield, a method capable of manufacturing high-purity D-psicose in high yield is required.

An embodiment of the present invention provides a method for manufacturing high-purity D-psicose in high yield.

According to an embodiment of the present invention, a method for manufacturing D-psicose includes: subjecting D-fructose to D-psicose epimerization to produce a solution containing D-psicose; The solution of psicose is subjected to a first cooling and ion purification; the purified solution of D-psicose is subjected to a first concentration and a second cooling; and the D-containing solution which has been subjected to the first concentration and the second cooling is D- The solution of psicose is subjected to chromatography to obtain a mother liquor containing D-fructose and a separated solution containing D-psicose; and the separated solution containing D-psicose is subjected to a second concentration and a third cooling to D-psicose crystals were obtained, in which the D-fructose-containing mother liquor produced by chromatography was reused in the D-psicose epimerization.

According to another embodiment of the present invention, a method for manufacturing D-psicose includes: subjecting D-fructose to D-psicose epimerization to produce a solution containing D-psicose; -The solution of psicose is subjected to a first cooling and ion purification; the purified solution of D-psicose is subjected to a first concentration and a second cooling; the solution containing D which has been subjected to the first concentration and the second cooling is D -The solution of psicose is subjected to chromatography to obtain a mother liquor containing D-fructose and a separated solution containing D-psicose; and the separation solution containing D-psicose is subjected to a second concentration and a third cooling To obtain D-psicose crystals, the D-fructose-containing mother liquor produced by chromatography is used in the D-psicose epimerization, and the first cooling and ion purification, and the first concentration In either of the second cooling and the chromatography, the mother liquor generated from the D-psicose crystals is reused.

In one embodiment or another embodiment, the method may further comprise, before reusing the mother liquor containing D-fructose produced by chromatography or the mother liquor produced by crystallizing D-psicose, The mother liquor containing D-fructose or the mother liquor derived from D-psicose crystals is cooled to 25 ° C to 45 ° C (especially 30 ° C to 40 ° C). The chromatography may be, for example, continuous chromatography.

In an embodiment or another embodiment, the first cooling and the third cooling may represent a process of reducing the solution temperature or the ambient temperature to 25 ° C to 45 ° C (especially 30 ° C to 40 ° C), and the second cooling may refer to The process of reducing the solution temperature or the ambient temperature to 45 ° C to 65 ° C (especially 50 ° C to 60 ° C).

In one embodiment or another embodiment, the first concentration refers to condensing the purified solution containing D-psicose to D-psicone having a brix of 50 to 70. Sugar concentration process. The second concentration refers to a process of concentrating D-psicose in the separation solution to a concentration of 75 Brid or more (for example, 80 Brid or more).

In one embodiment or another embodiment, as the D-fructose-containing mother liquor produced by chromatography, a D-fructose-containing mother liquor having a purity of 70% (w / w) or more can be used. Dissolved. As the mother liquor produced from the crystal of D-psicose, a D-psicose-containing dissolved fraction having a purity of 90% (w / w) or more can be used.

In one embodiment or another embodiment, the D-psicose epimerization may include a temperature of 40 ° C to 70 ° C (especially 40 ° C to 60 ° C, more specifically 40 ° C to 50 ° C) at D-psicose epimerase, a variant of the D-psicose epimerase, a strain capable of producing the D-psicose epimerase, or a culture of the strain in the presence of Table isomerization.

In one embodiment or another embodiment, ion purification may be performed using a strongly acidic cation exchange resin and / or a weakly basic anion exchange resin.

In one embodiment or another, the D-psicose-containing separation solution may contain 93% (w / w) or more than 93% (w / w) (especially 95% (w / w) or more than 95%). % (w / w)).

In one embodiment or another embodiment, the first to third cooling may be heat exchange cooling. Advantageously, due to the turbulence of the fluid, the heat exchange cooling provides a wider heat transfer area and faster heat exchange, prevents scale formation to reduce the function reduction, thereby providing higher economic feasibility.

In an embodiment or another embodiment, the D-psicose crystals obtained may have 95% (w / w) or greater than 95% (w / w) (more particularly 99% (w / w) or Greater than 99% (w / w)) purity.

In one embodiment or another embodiment, the D-psicose crystals prepared by the method as described above may have a yield of 75% or more, especially 85% or more.

In another embodiment, the method for manufacturing D-psicose provides D-psicose having a purity of 99% (w / w) or more.

The method for manufacturing D-psicose according to the embodiment of the present invention can prevent D-psicose from thermal deformation and enable D-psicose to be manufactured in a high yield through a stable process.

In addition, the method for manufacturing D-psicose according to an embodiment of the present invention collects a mother liquor containing D-fructose produced by chromatography and / or a mother liquor produced from D-psicose crystals, which are transferred to D -Epoxose sugar isomerization, first cooling and ion purification, first concentration and reuse in cooling or chromatography, so as to promote the stability of D-psicose through chromatography after long-term recycling Isolate and ensure the production of D-psicose in high yields.

The above and other aspects, features, and advantages of the present invention will become apparent from the detailed description of the following embodiments in conjunction with the accompanying drawings, wherein: FIG. 1 depicts D-psicone according to an embodiment of the present invention Schematic diagram of sugar production method.

FIG. 2 is a schematic diagram depicting a method for manufacturing D-psicose according to another embodiment of the present invention.

FIG. 3 is a schematic diagram depicting a method for producing D-psicose according to Comparative Example 2. FIG.

Figures 4 to 8 show graphs depicting the purity% (w / w) of D-psicose depending on the temperature and concentration of D-psicose.

Hereinafter, embodiments of the present invention will be described in more detail. A description of details apparent to those skilled in the art having ordinary knowledge in this technical field or related fields will be omitted herein.

In one embodiment of the present invention, a method for producing D-psicose includes: subjecting D-fructose to D-psicose epimerization to produce a solution containing D-psicose; -The solution of psicose is subjected to a first cooling and ion purification; the purified solution of D-psicose is subjected to a first concentration and a second cooling; the solution containing D which has been subjected to the first concentration and the second cooling is D -The solution of psicose is subjected to chromatography to obtain a mother liquor containing D-fructose and a separated solution containing D-psicose; and the separation solution containing D-psicose is subjected to a second concentration and a third cooling To obtain D-psicose crystals, the D-fructose-containing mother liquor produced by chromatography is used in the D-psicose epimerization.

In another embodiment of the present invention, a method for manufacturing D-psicose includes: subjecting D-fructose to D-psicose epimerization to produce a solution containing D-psicose; The solution of D-psicose was subjected to first cooling and ion purification; the purified solution of D-psicose was subjected to first concentration and second cooling; The D-psicose solution was subjected to chromatography to obtain a mother liquor containing D-fructose and a separation solution containing D-psicose; subjecting the separation solution containing D-psicose to a second concentration and a third cooling To obtain D-psicose crystals, the D-fructose-containing mother liquor produced by chromatography is used in the D-psicose epimerization, and the first cooling and ion purification, and the first concentration In either of the second cooling and the chromatography, the mother liquor produced by D-psicose crystallization is reused.

In the method according to the present invention, the product produced by chromatography is reused in any one of D-psicose epimerization, first cooling and ion purification, first concentration and second cooling, and chromatography. Contains D-fructose mother liquor or mother liquor produced through crystallization of D-psicose, and thus D-psicose can be manufactured with high yield, thereby improving manufacturing efficiency while reducing manufacturing costs.

First, D-fructose used as epimerized substrate can be used at a concentration of 30 Brid (%) to 50 Brid (%), and it can be dissolved at a temperature of 30 ° C to 40 ° C during use Under the water. Alternatively, D-fructose may be mixed with the mother liquor containing D-fructose produced by chromatography and at a temperature of 30 ° C to 40 ° C at a concentration of 30 Brid (%) to 50 Brid (%) use. Herein, as the D-fructose-containing mother liquor produced by chromatography, a purity of 70% (w / w) or more than 70% (w / w) (particularly 75% (w / w) or more than 75) can be used. % (w / w)) containing D-fructose soluble fraction.

D-psicose epimerization refers to D-psicose epimerase, a variant of the D-psicose epimerase, capable of producing the D-psicose A process for producing D-psicose through epimerization of D-fructose in the presence of a strain of epi-isomerase enzyme or culture. The D-psicose epimerase according to the present invention may include microorganisms derived from various donors such as Agrobacterium tumefaciens , Flavonifractor plauti , and Clostridium hylemonae ) enzyme or a variant thereof. The strain used for transformation may include, but is not limited to, Escherichia coli or genus Corynebacterium , genus Bacillus , and genus Aspergillus . For example, a strain transformed with E. coli may include BL21 (DE3) / pET24-ATPE [Korean Patent Publication No. 10-2011-0035805A], BL21 (DE3) / pET24-ATPE-2 [Korean Patent No. 10 -1203856]. Corynebacterium strains can include Corynebacterium glutamicum ATCC13032 / pCJ-1-ATPE [KCCM11046 disclosed in Korean Patent Publication No. 10-2011-0035805A], glutamic acid stick Bacillus sp.ATCC13032 / pFIS-1-ATPE-2 [Deposit number KCCM11204P disclosed in Korean Patent No. 10-1203856], Corynebacterium glutamicum CJ KY [Deposition number disclosed in Korean Patent No. 10-1455759 KCCM11403P], Corynebacterium glutamicum ATCC13032 / pFIS-2-ATPE-2 [KCCM11678P disclosed in Korean Patent Publication No. 10-2015-0047111A], and the like.

In one embodiment, D-psicose epimerase, a variant of said D-psicose epimerase, can be produced by immobilizing a carrier such as sodium alginate. Strains of enzymes or cultures of ptoseose epimerase, areomerized equipment (e.g., column) is filled with immobilized enzyme, and a solution containing D-fructose is supplied to the packed column for epimerization . For epimerization, the temperature in the equipment can be maintained at 40 ° C to 70 ° C, such as 40 ° C to 55 ° C. The temperature of the supplied D-fructose-containing solution can be increased, for example, via a heat exchanger at a heating rate of 5 ° C to 20 ° C per hour to 40 ° C to 60 ° C (for example, 50 ° C) so that the SV (space velocity: flow rate) (L) / hour (Hr) / resin amount (L)) becomes 0.5 to 3. D-psicose produced by epimerization may have a purity of about 15% (w / w) to about 35% (w / w), such as about 20% (w / w) to about 30% (w / w).

The D-psicose-containing solution produced above was subjected to a first cooling and ion purification. The first cooling refers to a process of reducing the solution temperature or the ambient temperature to 25 ° C to 45 ° C (especially 30 ° C to 40 ° C). In one embodiment, the mother liquor produced by D-psicose crystals can be reused in the first cooling and ion purification. Specifically, it can be cold However, the temperature is slowly decreased through the heat exchanger at a rate of 1 ° C to 10 ° C per hour. If D-psicose is continuously exposed to a certain temperature, D-psicose can be decomposed through thermal deformation, thereby reducing the purity of D-psicose in the manufacturing process. Therefore, it is difficult to achieve high-yield production of D-psicose (see FIGS. 4 to 8). Therefore, according to the present invention, thermal deformation of D-psicose is prevented by controlling the temperature in each process of the method.

Subsequently, ion purification refers to a purification process that allows a cooled solution to pass through a column packed with a strongly acidic cation exchange resin and / or a weakly basic anion exchange resin. If a strongly basic anionic resin is used, D-psicose may be deformed at a low temperature of 25 ° C to 45 ° C, resulting in a decrease in purity. Therefore, in order to produce D-psicose having a high yield, a strongly acidic cation exchange resin or a weakly basic anion exchange resin may be used. More specifically, a 100% weakly basic anion exchange resin was used. The cation exchange resin and the anion exchange resin can be used simultaneously to effectively remove the ionic components. In this case, the ratio of the cation exchange resin to the anion exchange resin may be 1: 0.5 to 1: 3. During the ion purification, a temperature of 25 ° C to 45 ° C (especially 30 ° C to 40 ° C) is maintained to prevent D-psicose from deforming. Therefore, it is possible to remove the ionic components contained as a contaminant in the D-psicose-containing solution. After ion purification, the content of ionic components is 20 microsiemens per centimeter (microsiemens) or less than 20 microsiemens per centimeter (especially 10 microsiemens per centimeter or less than 10 microsiemens per centimeter), as measured by a conductivity meter . D-psicose in the ion purification solution has a purity of about 10% (w / w) to about 35% (w / w).

Subsequently, the ion-purified D-psicose-containing solution was subjected to a first concentration and a second cooling.

The first concentration refers to concentrating the ion-purified D-psicose-containing solution so that A process for obtaining the solution having a D-psicose concentration of 50 to 70 brids (for example, 55 to 65 brids). Specifically, it is possible to concentrate the ion-purified D-psicose-containing solution at 60 ° C to 80 ° C (for example, at a temperature of 65 ° C to 75 ° C) for 1 minute to 1 hour (especially 1 minute to 30). minute). For concentration, a low temperature evaporator can be used to prevent D-psicose from deforming. The term Brix, as used herein, refers to the weight percentage of D-psicose or D-fructose based on the total weight of the solution. After the first concentration, a second cooling may be performed. When the second cooling is performed, the temperature of the solution or the environment can be lowered to a temperature at least 10 ° C lower than the temperature of the first concentration. In one embodiment, the temperature of the second cooling may be 50 ° C to 60 ° C. Specifically, the temperature may be slowly decreased through the heat exchanger at a cooling rate of 5 ° C to 25 ° C per hour. In one embodiment, the mother liquor produced by D-psicose crystals can be reused in the first concentration and / or the second cooling.

Thereafter, the concentrated and cooled D-psicose-containing solution may be subjected to chromatography to obtain a mother liquor and a separated solution containing D-psicose.

Chromatography refers to the process of separating D-psicose using the weaker bonding strength between D-psicose and metal ions connected to the ionic resin. For example, chromatography can be continuous chromatography. The ionic resin used in chromatography can be a strongly acidic cation exchange resin linked to K, Na, Ca or Mg residues. In view of the separation of D-psicose and D-fructose, K, Ca or Na should be used. By chromatography, it is possible to obtain a D-fructose-containing solution and a D-psicose-containing separation solution. The separation solution containing D-psicose can be 90% (w / w) or greater than 90% (w / w) (e.g., 95% (w / w) or greater than 95% (w / w)). Contains D-psicose-soluble fractions. Specifically, D-psicose has a purity of 90% (w / w) to 99% (w / w) or greater than 99% (w / w).

D-fructose-containing solutions can be 70% (w / w) or greater (w / w) D-fructose-containing dissolved fraction. D-fructose-containing solutions can be reused in the epimerization of D-psicose. Before the reuse, the method may further include cooling the solution to a temperature of 25 ° C to 45 ° C (especially 30 ° C to 40 ° C).

Thereafter, the separation solution containing D-psicose was subjected to a second concentration and a third cooling to obtain D-psicose crystals. Make D containing 90% (w / w) or greater than 90% (w / w) purity (e.g. 95% (w / w) or greater than 95% (w / w) purity) D-containing -The solution of psicose is subjected to a second concentration such that the separation solution has a D-psicose concentration of 75 brids or more (e.g. 80 brids or more). Concentration is carried out especially at 60 ° C to 80 ° C (for example 65 ° C to 75 ° C) for 1 minute to 1 hour (especially 1 minute to 30 minutes). For concentration, it is possible to use a low temperature evaporator to prevent D-psicose from deforming. After the second concentration, a third cooling may be performed. The third cooling is used to crystallize D-psicose, and may include reducing the solution temperature or the ambient temperature to 30 ° C to 40 ° C. More specifically, the third cooling may be performed slowly so that the solution temperature or the ambient temperature decreases by 5 ° C to 20 ° C up to 25 ° C to 45 ° C (for example, 30 ° C to 40 ° C) per hour. Within the temperature range, heating and cooling may be repeated 5 to 10 times for 40 to 120 hours for crystallization. The obtained D-psicose crystals can be further dehydrated and dried. The D-psicose crystals obtained may have a purity of 95% or more, especially 99% or more. In addition, D-psicose crystals may have a yield of 75% or more, 80% or more, or 85% or more, as calculated according to Equation 1: Yield (%) = (Weight of dehydrated and dried D-psicose crystals / weight of D-psicose to be crystallized contained in solution) × 100.

When calculating the yield, Liquid chromatography (HPLC) analysis determined the weight (g / liter) of D-psicose to be crystallized contained in the solution. Subsequently, the measured value is substituted into Equation 1 to calculate the weight of D-psicose contained in a specific solution (liter).

Mother liquors other than the D-psicose crystals produced after the D-psicose crystals can be reused in each of the above processes. In one embodiment, the mother liquor produced after crystallization can be reused in one or more processes selected from the first cooling and ion purification, the first concentration and second cooling, and the chromatography process. The mother liquor produced from D-psicose crystals can be cooled to 25 ° C to 45 ° C and then reused. The mother liquor produced from D-psicose crystals may have a purity of 90% (w / w) or greater than 90% (w / w), a purity of 92% (w / w) or greater than 92% (w / w) , Or a D-psicose-containing dissolved fraction having a purity of 95% (w / w) or greater than 95% (w / w).

Hereinafter, the present invention will be described in more detail with reference to some examples. It should be understood that these examples are provided for illustration only and should not be construed as limiting the invention in any way.

This article will omit detailed descriptions obvious to those with ordinary knowledge in the field.

Examples

Example 1

Mix crystalline fructose with purity of 99% (w / w) or greater than 99% (w / w) in the solution tank, mother liquor collected from continuous chromatography at about 30 ° C, and mother liquor collected from crystallization at about 30 ° C And water at about 30 ° C to prepare a 50 Brix (%) solution of the substrate for the enzyme reaction.

As disclosed in Korean Patent Application No. 10-2009-0118465, by immobilizing from Corynebacterium glutamicum KCCM 11046 points on sodium alginate as a carrier The D-psicose epimerase is subjected to an enzymatic reaction, and then an isomerization device (isomerization tower, manufactured by Hanju Machine Industry Inc.) is filled with an immobilized enzyme, and the prepared The enzyme reaction substrate solution is applied to the isomerization tower and the substrate solution is heated to 50 ° C at 5 ° C to 20 ° C per hour through a heat exchanger so that SV (space velocity: flow rate (L) / hour (Hr) ) / Resin amount (L)) becomes 0.5. Here, the obtained D-psicose has a purity of about 24% (w / w).

The D-psicose solution having a purity of 24% (w / w) is subjected to a first cooling, which is reduced to a temperature of 30 ° C to 40 ° C at a rate of 5 ° C to 10 ° C per hour through a heat exchanger, and then Through a column filled with a strongly acidic cation exchange resin substituted with a hydrogen group (Lewatit S 1668) and a column filled with a weakly basic anion exchange resin substituted with a hydroxyl group (Lutet S 4528) So that SV (space velocity: flow rate (L) / hour (Hr) / resin amount (L)) becomes 3, thereby removing residual ionic components in the enzyme reaction solution. Removal of ionic components was measured using a conductive meter. Here, the conductivity is adjusted to not more than 10 micro-Siemens per cm, and the purity of D-psicose is maintained at 24% (w / w).

The ion-purified D-psicose-containing solution was introduced into a low-temperature evaporator (Forced Thin Film Evaporator) at 65 ° C to 75 ° C, Welcronhantec Co., Ltd. )), Concentrated to 60 Brid (%) (D-psicose solution × 100 / total solution) for a short time of 10 minutes to 15 minutes. The resulting solution was subjected to a second cooling via a heat exchanger at a rate of 5 ° C to 25 ° C per hour, and the resulting solution was then passed at 50 ° C to 60 ° C by being filled with a strongly acidic cation exchange resin (having calcium activity attached thereto). Group). Through such continuous chromatography, D-psicose-containing soluble fractions with a purity of 95% (w / w) or greater are separated from 75% (w / w) Or D-fructose-containing dissolved fractions with a purity of greater than 75% (w / w).

Collect the mother liquor separated by continuous chromatography, that is, D-fructose with a purity of 75% (w / w) or more, and dissolve it at a rate of 20 ° C to 30 ° C per hour . When the dissociation reaches 30 ° C., the dissociation is recycled to the enzyme reaction process.

D-Allow with a purity of 95% (w / w) or greater than 95% (w / w) will be separated by continuous chromatography at 65 ° C to 75 ° C and within a short period of 10 minutes to 15 minutes The ketose solution was concentrated to 80 Brix (%). Quickly cool the concentrated D-psicose solution with a purity of 95% (w / w) or greater than 95% (w / w) through a heat exchanger at a rate of 5 ° C to 20 ° C per hour . Heating and cooling were repeated 5 to 10 times in a temperature range of 35 ° C to 40 ° C, so that the crystallization was performed for 80 hours to 120 hours, thereby obtaining D-psicose (see Fig. 1). In addition, the mother liquor separated by crystallization, that is, a dissolution fraction of D-psicose having a purity of 90% or more, was collected, and cooled to 30 ° C and filled with a strong acid substituted with a hydrogen group. A column of a basic cation exchange resin and a column filled with a weakly basic anion exchange resin substituted with a hydroxyl group.

Through the above procedure, the D-psicose solution is concentrated in a low-temperature evaporator for a short period of time, and the process temperature is adjusted to a low temperature through a heat exchanger, so that an excellent yield of 75% or more is obtained with a 99% High-purity D-psicose with a purity of% or greater than 99%.

Comparative Example 1

A high-purity D-psicose sugar having a purity of 99% or more was produced by the same method as in Example 1 except that the cooling process including the first cooling and the second cooling was omitted, wherein Ketose epimerization The D-fructose-containing mother liquor was produced in the same, and the mother liquor produced by D-psicose crystallization was reused. The manufacturing yield was 60%.

The purity of D-psicose decreases at a certain concentration or lower and after a certain period of time (see Figs. 4 to 8). Therefore, if the D-fructose mother liquor separated by continuous chromatography and the mother liquor separated by crystallization are continuously recycled without cooling, the purity of D-psicose introduced into continuous chromatography will be compared to 24% ( w / w) The initial purity is reduced in proportion to the number of repetitions of the recycle, and the high purity D-psicose having a purity of 95% (w / w) or greater than 95% (w / w) to be crystallized The yield of dissociated fractions will also decrease. Therefore, such lower yields of high-purity D-psicose dissolving fractions that can be considered to have a purity of 95% (w / w) or greater than 95% (w / w) serve as having 99% or greater The reason why the production yield of high-purity D-psicose with a purity of% is reduced. In addition, in the process of continuous chromatographic separation, the deformed impurities derived from D-psicose are separated into D-fructose dissolved fractions, which causes problems in continuous chromatographic separation as the number of recycles accumulates. In order to control the accumulation of such impurities, about 5% (v / v) to 10% (v / v) of D-fructose dissolution is discarded, thereby causing high-purity D-alo with a purity of 99% or more The production yield of ketose decreases.

If about 5% (v / v) to 10% (v / v) of the D-fructose mother liquor separated by continuous chromatography is not discarded, the purity of D-psicose introduced into continuous chromatography will be Continuously decreases due to accumulated impurities. Therefore, in order to separate high-purity D-psicose having a purity of 95% (w / w) or more, the separation conditions of continuous chromatography must be continuously modified, resulting in industrial manufacturing efficiency. decline. In addition, in order to separate a high-purity D-psicose having a purity of 95% (w / w) or more, the amount of deionized water is also increased, thereby causing a burden on the concentration process. Therefore, a bottleneck phenomenon occurs in the process flow. As a result, the manufacturing volume is reduced and the manufacturing cost is increased.

Comparative Example 2

Except for omitting the reuse of mother liquid containing D-fructose produced by continuous chromatography in D-psicose epimerization and the reuse of mother liquid produced by D-psicose crystallization, 1 The same method produces D-psicose with high purity having a purity of 99% or more. The manufacturing yield was 20% (see Figure 3).

Although some embodiments have been described herein, those with ordinary knowledge in the art should understand that these embodiments are given by way of illustration only, and various modifications and changes can be made without departing from the spirit and scope of the present invention. And changes. Therefore, the scope of the invention should be limited only by the scope of the accompanying patent applications and their equivalents.

Claims (16)

A method for manufacturing D-psicose, comprising: subjecting D-fructose to isomerization of D-psicose to produce a D-psicose-containing solution, and causing the D-psicose-containing solution The solution is subjected to first cooling and ion purification, and the purified solution containing D-psicose is subjected to first concentration and second cooling, so that it has been subjected to the first concentration and the second cooling The D-psicose-containing solution is subjected to chromatography to obtain a D-fructose-containing mother liquor and a D-psicose-containing separation solution, and the D-psicose-containing separation solution is subjected to the first Second concentration and third cooling to obtain D-psicose crystals, wherein the D-fructose-containing mother liquor produced by chromatography is used in the D-psicose epimerization, wherein The method for manufacturing D-psicose further includes reuse of the D-arc in the first cooling and the ion purification, the first concentration and the second cooling, or the chromatography Mother liquor produced by crystallization of loxulose. The method for manufacturing D-psicose as described in item 1 of the scope of the patent application further includes: reusing the crystals produced by the D-psicose in the first cooling and the ion purification The mother liquor. The method for manufacturing D-psicose described in item 1 of the patent application scope, wherein the chromatography is continuous chromatography. The method for producing D-psicose according to any one of claims 1 to 3, wherein the mother liquid containing D-fructose produced by the chromatography is added to the D -The mother liquor, which is cooled to 25 ° C to 45 ° C before being reused in the crystallization of alloxan sugar, or produced by the crystallization of the D-aloxulose in the first cooling and the ion purification, The first concentration and the second cooling or cooling to 25 ° C to 45 ° C before reuse in the chromatography. The method for manufacturing D-psicose according to any one of claims 1 to 3, wherein the first cooling and the third cooling are performed by cooling the solution temperature or the ambient temperature to It is performed at 25 ° C to 45 ° C, and the second cooling is performed by lowering the temperature of the solution or the ambient temperature to 45 ° C to 65 ° C. The method for manufacturing D-psicose according to item 5 of the patent application scope, wherein the first cooling and the third cooling are performed by cooling the solution temperature or the ambient temperature to 30 ° C to 40 The temperature is carried out at 0 ° C., and the second cooling is performed by lowering the temperature of the solution or the ambient temperature to 50 ° C. to 60 ° C. The method for producing D-psicose according to any one of claims 1 to 3, wherein the first concentration is performed so that the purified D-psicone-containing The sugar solution has a D-psicose concentration of 50-70 degrees Brit, and the second concentration is performed so that the D-psicose-containing separated solution has a 75-degree Brit or greater The concentration of D-psicose at 75 Brido. The method for producing D-psicose according to any one of claims 1 to 3, wherein the mother liquid containing D-fructose produced by the chromatography has 70% ( w / w) or greater than 70% (w / w) purity D-fructose-containing dissociated fraction. The method for manufacturing D-psicose as described in item 1 of the patent application range, wherein the mother liquor produced by crystallization of the D-psicose is 90% (w / w) or greater than 90% (w / w) purity of D-psicose-containing dissociated fraction. The method for producing D-psicose according to any one of the first to third items of the patent application scope, wherein the D-psicose isomerization is at 40 ° C to 70 ° C and at The presence of a D-psicose epimerase, a variant of the D-psicose epimerase, an enzyme or a culture capable of producing the D-psicose epimerase Proceed. The method for producing D-psicose according to any one of the first to third patent claims, wherein the ion purification uses a strongly acidic cation exchange resin or a weakly basic anion exchange resin. The method for manufacturing D-psicose as described in any one of the first to third items of the patent application range, wherein the separation solution containing D-psicose contains 93% (w / w) Or D-psicose with purity greater than 93% (w / w). The method for manufacturing D-psicose according to any one of the first to third patent claims, wherein any one or more of the first cooling to the third cooling is heat exchange cool down. The method for manufacturing D-psicose according to any one of the first to third items of the patent application range, wherein the obtained D-psicose crystals have a purity of 95% or greater than 95% . The method for manufacturing D-psicose according to item 14 of the patent application scope, wherein the obtained D-psicose crystals have a purity of 99% or greater than 99%. The method for manufacturing D-psicose according to any one of claims 1 to 3, wherein the D-psicose crystals have a yield of 75% or more.